THE DEVELOPMENT OF PROTEIN VANILLA ICE CREAM FORMULATED WITH

STEVIA AS A SWEETENER

SARANKORN PORNPETCHLERK

A SENIOR PROJECT SUBMITTED IN PARTIAL FULFILMENT OF THE

REQUIREMENTS FOR THE DEGREE OF BACHELOR OF SCIENCE (FOOD
SCIENCE AND TECHNOLOGY)

MAHIDOL UNIVERSITY INTERNATIONAL COLLEGE

MAHIDOL UNIVERSITY
2019

COPYRIGHT OF MAHIDOL UNIVERSITY

THE DEVELOPMENT OF PROTEIN VANILLA ICE CREAM FORMULATED WITH
STEVIA AS A SWEETENER

was submitted to the Mahidol University International College, Mahidol University

for the degree of Bachelor of Science (Food Science and Technology)
on
August 29, 2019

……………………………….…
Sarankorn Pornpetchlerk
Candidate

……………………………….…
Dr. Varongsiri Kemsawasd
Advisor

……………………………….…
 ACKNOWLEDGEMENTS

Four years in university have been a tough, exhausting, but still memorable and beautiful

journey for me. This senior project would not have been completed without the support from my

family, friends, teachers and laboratory staff.

First of all, I would like to thank Dr. Varongsiri Kemsawasd. I would not be able to do the

thesis without your help. You have been very supportive and helpful. You are one of my best

teachers on and off campus. I always feel grateful for your suggestions and advice you have

given to me.

I also would like to dedicate this acknowledgements to laboratory staffs, especially Miss

Supaporn Phomsurin, for being another great supporter, not just for the thesis but for all the

classes I have taken during my four years in this university.

Lastly, I would like to thank all my friends for being together for four years. It would

have been a lot tougher and more nerve-wracking without you all in my life.

Sarankorn Pornpetchlerk
THE DEVELOPMENT OF PROTEIN VANILLA ICE CREAM FORMULATED WITH
STEVIA AS A SWEETENER

SARANKORN PORNPETCHLERK 5880121

B.SC. (FOOD SCIENCE AND TECHNOLOGY)

SENIOR PROJECT ADVISOR: DR. VARONGSIRI KEMSAWASD

ABSTRACT

The low-fat vanilla ice cream fortified with whey protein concentrate was formulated

with an aim to produce ice cream for athletes or individuals who are physically active. Whey

protein can be found as a by-product from the cheese-making industry. It is often used in

exercise supplements due to the fact that whey protein contains all nine essential amino acids

that are necessary for several body functions. It is also proven to play an important role in weight

control. In this study, stevia is used to replace some of the sugar content. Stevia is typically

undigested by the human digestive tract and cannot be accumulated in the body; hence, the

energy provided by the stevia can be seen as zero calories. There were two formulations of ice

cream mix which were 25% whey substitution and 15% whey substitution. After the sensory

evaluation was performed by 30 healthy panelists, 15% whey substitution was selected as the

best formulation. The ice cream was composed of 15% whey protein substitution, half and half

cream, low-fat milk, sugar, stevia powder, water, and vanilla extract. There was a significant

increase in %overrun compared to the control sample, which did not contain whey protein

concentrate and formulated with full-fat milk. The whey protein ice cream also exhibited a

higher moisture content. Moreover, the melting rate was significantly reduced to 0.5252±0.005,
while the control sample showed the melting rate of 0.9069±0.006. This reduction in the melting

of the ice cream is caused by the increase of protein content and milk fat destabilisation.

KEYWORDS: WHEY PROTEIN/ STEVIA/ LOW-FAT/ ICE CREAM

 CHAPTER I

INTRODUCTION

Ice cream is a dairy product made from dairy ingredient mix which has been processed

through various steps. Ice cream is known as a complex food matrix as it contains multiple

components such as stabilizers, flavouring agents, sweeteners, emulsifiers with a dairy ingredient

as a major part. Frozen dairy food originated from ice beverages. Later on, the ice cream was

developed with the introduction of homogenizers, pasteurizers and the freezers into the industry.

Ice cream products have become more recognized and obtained higher market share due to the

increase in popularity, not just in children and young adults but also adults and the elders,

because the technological advancement has made the ice cream ubiquitously available to

consumers at very low cost.

Theoretically, ice cream is considered to be a nutritious food. It is an easily-made and

highly digestible food, so it is frequently substituted in hospital meal plan for patients who have

difficulties in swallowing. Ice cream can be classified as a nutrient-dense product depending on

its ingredients added. The ice cream, usually contains carbohydrates, proteins, fats and several

minerals which are the primary nutrients required daily by the human body to be able to function

properly.

Nowadays, people are more concerned about their health and diets. The trend of

exercising has been rising in every age. The personal nutrition requirement is varied depending

on the daily activities and lifestyles. The people, who are not physically active, will require

standard amounts of macronutrients. However, athletes or physically active people may require

special dietary needs, especially proteins. Protein is one of the essential macronutrients required
by the body as it plays a vital role in being building blocks for tissues, enzymes, and hormones.

It is also responsible for building muscles and repairing the muscle breakdown. The

recommended daily allowance for protein intake is 0.8 g per kilogram of body weight. However,

the requirement is relatively higher for athletes. Due to the higher demand in protein

consumption, whey protein is frequently supplied into the food product.

Whey protein is a protein part that can be obtained as a by-product from cheese-making.

It contains essential amino acids and low lactose content. Whey protein has been found to assist

in weight loss, reduce blood pressure, and decrease cholesterol level. There are three main forms

of whey which are whey protein powder, whey concentrate, and whey isolate. Each type contains

different amounts of proteins. The selection of protein supplements based on the purpose of

consumer’s consumption. For instance, individuals that are losing weight are more suitable with

whey isolate because it contains less amount of fats than whey concentrate. Even though, whey

protein is beneficial to human health, it still has some limitations. Whey protein is considered as

dairy product, therefore, it cannot be consumed by vegetarians. Moreover, whey protein also has

its distinct flavour and aroma, which is sometimes unpreferred by the consumers.

As aforementioned, the trend of being healthy is increasing significantly. The ingredients

added to the food have now become a major priority for the people who are concerned with their

health. The food industry is trying to cope with the change by finding the best substitution.

Sugars, short-chained, soluble carbohydrates e.g., sucrose, fructose, and glucose, provide the

sweet taste and serves as a bulking agent. However, high consumption of sugars can lead to

several diseases such as diabetes, cardiovascular diseases, and cancer. Consequently, sugars are

substituted with other types of sugars, for instance, sugar alcohol, aspartame, and stevia. Stevia,
the natural sweetener, is a sweetener derived from the leaves part of the Asteraceae family. It is

200 times sweeter than table sugar and does not give any calories. In contrast with commercial

sugars, stevia has been found to treat obesity, hypertension, and type II diabetes. In addition,

another factor concerned is fiber content in the food. Dietary fiber refers to the plant-based

nutrient that is indigestible in the intestinal tract. Fiber is essential for several body functions, for

instance, digestion, weight control, blood sugar regulation or food allergy prevention. Fiber

supplements are often taken or added with the food to aid people who are struggling to have an

appropriate amount of fiber. Inulin is a type of soluble fiber that is composed of fructose

molecules. Inulin serves as a prebiotic for gut bacteria. The research has proved that inulin can

improve the digestive system and control diabetes, as well as reduce constipation.
 CHAPTER II

OBJECTIVES

To formulate a protein-rich and low-fat ice cream product using whey protein concentrate

and stevia for athletes with the consumer acceptability.

 CHAPTER III

LITERATURE REVIEWS

Nutrition requirement

The term ‘athlete’ cover all the people who compete in any types of sports, for instance,

weight-lifting, team sports, and also sports aiming for endurance. To maximize the performance

on the field or in the gym requires several factors to be done, diets are one of the essential

components that can lead the athlete to their full potential. Nutritional intake heavily impacts the

body function of the athletes and can improve the overall performance if they are fulfilled by all

the important requirement. Macronutrients such as carbohydrates, protein, and fat are varied and

calculated according to the goal of the athletes. These macronutrients play different roles in the

body; therefore, the adequate requirement must be achieved daily to maintain the performance

quality. Carbohydrate intake is aimed to fuel for athletes who compete in endurance sports.

Consuming carbohydrates before exercise has shown to reduce the incidence of fatigue and

increase in performance improvement (Hawley et al., 1997). Recommendation for adequate

carbohydrate intake is approximately 3-5 g per 1 kg of body mass. However, the amount will be

increased to compensate for the higher energy used in daily activities (Burke et al., 2011).

Another essential macronutrient is fat. Fat is required to regulate many bodily functions such as

absorption of fat-soluble vitamins, providing energy for muscle metabolism, and also improve

brain health. Nevertheless, the fat consumed should be derived from good sources, which contain

high unsaturated fats and all the essential amino acids, for instance, salmons, avocadoes, and

nuts. The recommendation has stated that at least 20% of total calories consumed in a day should

come from good fats (Rodriguez et al., 2013). Lastly, proteins are considered as one of the most
important keys that contribute to the athlete’s full potential. Protein serves as the building blocks

for muscles and soft tissues. It also is required for hormones and enzymes synthesis. In general,

the dietary reference intake of protein is approximately 0.8g per kilogram of body weight for

sedentary individuals (Phillips, 2006). However, the requirement will be higher for athletes or

active people. For endurance and strength-trained athletes, protein intake should be around 1.2 or

1.7g per kilogram of body weight. Protein intake is increased to compensate with the protein

oxidation during the long training session. Moreover, it is also needed in higher amount for

strength athletes because protein is used to support muscle growth and maintain the muscle mass.

This DRI of protein can be achieved through regular diets, and however, if the intensity of the

training is higher, protein supplements may be required to prevent body mass loss sufficiently.

Ice cream

Ice cream is a mixture of milk fat, milk solids-non-fat, stabilisers, sweeteners,

emulsifiers, and water. The ingredients selected by the manufacturers are generally based on the

cost and availability of the materials. Each ingredient used in the processing has a significant

impact on the final product.

The milk fat mainly contributes to flavor and the texture of the ice cream. The fat present

in the mixture has been found to have the ability to react with the flavor compound which will

enhance the overall mouthfeel. The creaminess of the ice cream is also expected to be lower

when the fat content is reduced. Moreover, fat can also alter the vapor pressure of the flavor

compound by trapping the lipophilic volatiles in the lipid compounds found in food. As a

consequence, the headspace concentration of volatile compounds is relatively lower than the

low-fat food. (Keller et al., 1994). Milk solids-non-fat refers to lactose, casein, whey proteins,
minerals, vitamins, and other components of milk. The milk solids-non-fat includes skim milk

powder, condensed milk and whey protein concentrate ingredients. The protein from the milk

also plays an important role in developing the texture of ice cream.The viscosity of the ice cream

is usually enhanced due to the water-holding capacity of the proteins. This property prolongs the

meltdown time of ice cream and also helps to prevent the undesirable crystallisation, which leads

to the reduced iciness.

Sweeteners are responsible for the palatability of the ice cream. It is added to provide a

sweet taste, enhance the flavor, and also to decrease the freezing point of the mixture. Amount of

sweeteners added must be calculated in order to achieve the desired amount of total solids in the

ice cream. Sucrose is the most common sweetening agent used. However, glucose syrup can be

substituted for more smoothness and more chewy texture (Deosarkar et al., 2016). In this

health-oriented era, non-nutritive sweeteners can be used for the specific group of consumers, for

instance, diabetic patients or people who are losing weight because it does not give any calories

and has no effects on the blood sugar level.

Stabilizers are required in the ice cream processing to help stabilise the structure of ice

cream. Commonly, polysaccharides such as guar gum, locust bean gum, and xanthan are used as

a stabilizer. This component enhances the flavour suspension, provides smoothness to the final

texture and also increases the overall viscosity. Stabilizers are commonly used at 0.1%-0.5%;

however, low-fat ice cream might require higher amount of stabilizing agent (Deosarkar et al.,

2016).

Apart from stabilizers, emulsifiers such as mono- and diglycerides are also used to help

stabilize the mix of ice cream. Even though, the role of the stabilisers and emulsifiers seems to
be similar, it is different. Emulsifiers are added to control fat destabilization, reduce whipping

time and also increase the melting point of the ice cream

Ice cream production

For the preparation process, pasteurization is implemented to destroy pathogenic bacteria

in raw milk, reconstituted milk of ice cream mix. In this step, the product is heated at 69°C for at

least 30 minutes or 80°C for at least 25 seconds. After pasteurization, the ice cream mix is now

passed to the homogenization process. In this step, the mix is homogenized at high pressures.

Homogenization reduces the interfacial free energy of the fat globules, as a consequence, the fats

will have less chance to aggregate together and form clumps. Homogenization is mainly aimed

to produce uniform fat droplet size, resulting in a more uniform final product with smoother

texture.

After the mix has been homogenized, it will be aged for at least 4 hours at 2-4°C. Aging

is the process that allows hydration of milk proteins and stabilizers, crystallization of the fat

globules, and a membrane rearrangement to produce a smoother texture. In this stage, milk

proteins from the fat surface will be replaced by emulsifiers, which will improve the whipping

qualities of the mix.

The last step in the production line is the freezing step. During freezing, the heat in the

substance is removed, therefore, the equilibrium between water and ice is altered. The freezing

process is done in two stages. Firstly, the mix is passed through the heat exchanger under high

shear conditions to exhibit ice crystal nucleation and also to corporate air into the mix. Secondly,

freezing the packaged ice cream rapidly and forming the small ice crystals. For ice cream

production, freezing is one of the vital steps that can determine the quality of the final product.
During the incorporation of air, the foam is created. It leads to the partial destabilisation of fat

emulsion. The goal of the ice cream manufacturers is to produce the ice cream with ice crystals

that cannot be detected by the consumers. So, the freezing process must be strictly controlled.

The final ice cream product after freezing step must be at -5°C or lower.

Whey Protein

Whey protein is the liquid part of milk that is obtained as a by-product from the

cheese-making industry. Whey protein is consumed by many individuals due to its beneficial

nutrition. It is proven to maintain lean body mass while reducing the fat mass of the athletic

people because whey protein can increase muscle synthesis in response to exercise (Candow et

al., 2006). Whey protein is comprised of several proteins such as β-lactoglobulin (β-LG),

α-lactalbumin (α-LA), the heavy-and light-chain immunoglobulins (Igs), bovine serum albumin

(BSA), lactoferrin (LF), lactoperoxidase, and glycomacropeptide (GMP) (Wit, 1998). Whey

protein also contains 20 amino acids with all nine essential amino acids that take parts in

important body functions such as regulation of whole body protein metabolism, tissue growth

and repair, and also promote weight control.

Three main types of whey protein, as a result from different processing techniques, are

whey protein powder, whey protein concentrate, and whey protein isolate. Whey protein powder

is found as an additive in dairy, bakery, confectionery and snack products. It can be used in infant

formulas as well. For whey protein concentrate, it is obtained by removing water, lactose, ash

and some minerals from the whey powder through the process of ultrafiltration. It contains

higher biologically active components and proteins than whey isolate, which make it a better

supplement for the athletes. Lastly, whey protein isolate. It is the purest source of proteins. The
isolates contain more than 90% of protein concentration which fat and lactose are removed,

therefore, it is more suitable for lactose-intolerant and losing weight individuals. Even though; it

has the highest concentration of protein, it also contains some denatured proteins which are a

result from the manufacturing process. The denatured proteins loss the peptide bonds, and the

structure is altered, which leads to the reduction of the protein effectiveness (Hoffman & Falvo,

2004).

Table 1 : Composition (%) of whey protein forms (Geiser, 2003)

Component Whey Powder Whey Concentrate Whey Isolate

Protein 11-14.5 25-89 90+

Lactose 63-75 10-55 0.5

Milk Fat 1-1.5 2-10 0.5

Regardless of the forms, nowadays, whey protein is supplemented in many products due

to its health benefits and specific functional properties. Whey protein has an ability to form gels

which are capable of holding water, lipids, and other components, which leads to an

improvement in texture for many foods such as processed meat, dairy and bakery products

(Mullvihill & Kinsella, 1987).

Whey proteins are also added into the milk used for acid-fermented drinks. The addition

of whey protein causes the change of casein:whey protein ratio. It has a great impact on the

thermal stability of the milk (Kalab et al., 1983). However, if whey protein is added more than

1-2%, it could lead to the undesirable odour of the product.

 Whey proteins are not only beneficial to the food industry, but it has also been used in the

medical field frequently. In whey proteins, they contain glycoprotein called Lactoferrin.

Lactoferrin has antifungal, antibacterial, and antiviral properties. It is able to release the outer

membrane of gram-negative bacteria, therefore, it can also work as an antibiotic (Shah, 2000).

Moreover, lactoferrin in whey can exhibit anti-inflammatory properties by regulating the tumor

necrosis factor and interleukin 6, which will reduce the inflammation in the body.

From research, amino acids found in whey protein can increase glutathione concentration

in the tissues, enhance human immunity and also be able to detoxify potential carcinogens

(Bounous, 2000). Many researchers have stated that the whey has anticancer property because it

can prevent iron from causing oxidative damage to tissues due to its iron-binding capacity

(Weinberg, 1999).

Whey proteins have been one of the most influential products for exercise and weight

loss industry. It contains a high amount of branched chain amino acid or BCAA which serve as a

substrate for protein synthesis. Leucine found in BCAA is thought to enhance muscle

hypertrophy and increase muscle strength (Ha & Zemel, 2003). Glutathione in whey protein also

helps to reduce muscle inflammation after intense training (Viguie et al., 1988). Whey proteins

can also cope with obesity due to the fact that it contains calcium. Calcium in dairy products can

affect energy metabolism by regulating adipocyte lipid metabolism and triglyceride storage

(Zemel, 2003). Whey has been found in many fields due to its beneficial properties. To date,

there are no severe adverse effects reported. However, some forms of whey protein might not be

suitable for lactose-intolerant individuals.

Stevia
 Stevia is a sweetener that gives no calories to the consumer. It is naturally available.

Stevia, nowadays, is used to replace sugar in the food industry. The plant is originally cultivated

in South America. It has been used as a flavoring agent for more than 200 years by native people.

The chemical component that is responsible for sweetness in stevia is ‘steviol glycosides’. The

steviol glycosides that are most abundant are rebaudioside A and stevioside, which 200-400

times sweeter than sucrose (Ashwell, 2015). Steviol glycosides are undigested by human

gastrointestinal tract, but the gut bacteria hydrolyzes it into steviol. The steviol is later on

absorbed by the vein and metabolized by the liver, resulting in steviol glucuronide, which will be

excreted in the urine. The research has stated that stevia is not accumulated in the body,

therefore, the energy provided by the stevia can be assumed as zero calories (Gardana et al.,

2003). Thus, stevia extracts are often used as a food additive for weight loss individuals since it

does not contribute to calorie intake. Moreover, stevia is listed as generally recognized as safe

food or GRAS by US Food and Drug Administration. However, the Acceptable Daily Intake

(ADI) of high purity stevia extract is approximately 12 mg/kg of body weight per day. Stevia can

be used as a tool to reduce the energy intake, which has the potential to prevent obesity in the

patients.
 CHAPTER IV

MATERIALS AND METHODS

4.1 Materials

The materials used in this experiment were as follows; glass bowls, spatula, spoons,

water cups, disposable plastic cups, disposable plastic spoons, and 100 ml beakers.

The ingredients used in this study were as follows; Thermax whey proteins, Stevia extract

powder, Half and half cream, low-fat milk, milk, sugar, water and vanilla extract.

The instruments used are mainly as follows; Blender, Ice cream maker, Precision balance

(Aczet CY2202C), Moisture analyser (Mettler Toledo HB43-S), Calorimeter (Konica Minolta

CR-400 Chroma Meter), pH meter (HANNA HI 98107).

4.2 Methods

4.2.1 Ice cream process

The ice cream samples were prepared by weighing the ingredients precisely using the

precision balance. The major ingredients of the ice cream were

Whey proteins ice are mixed with half and half cream, milk, and water. In this study, the

different amount of stevia and whey proteins are varied. The ingredients are weighed by using

the weighing machine with 2 decimal places. To get a more uniform mix of ice cream, these

main components are blended together in the blender. Then, the mix was added with sugar and

stevia. Stevia was calculated to substitute 15% of sugars, as it provides really low calories and

does not affect blood sugar level. However, high amount of stevia can produce a bitter taste and

some consumers can detect the after taste, thus, it is not able to replace all of the sugar content.
Then, 1 teaspoon of vanilla extract was dropped into the mixture and put into the ice cream

maker for 40 minutes or until set. The ice cream is stored at -20 prior to further study.

4.2.2 Ice cream formulation

Different concentrations of stevia, low-fat milk, and whey proteins were used in order to

formulate the most healthy ice cream. The selected formulation of the ice cream was then used to

be further analyzed for physicochemical properties including overrun, melting rate, color, pH

and total solid content (degree Brix). The determination will be performed in duplicates.

4.2.3 Ice cream analysis

4.2.3.1 % Overrun

The overrun measurements were taken utilizing the weight of the mix in a fixed

volume container (500 ml beaker) and the weight of the fixed volume of ice cream (500

ml beaker) (Warren & Hartel, 2018). % Overrun was calculated as follows:

% Overrun = (Vol. of ice cream - Vol. of mix used)/Vol. of mix used x 100%

4.2.3.2 Melting Rate

One portion of ice cream was separated and placed on a mesh with a size of 0.3

cm. The mesh was put on top of the beaker. The sample was placed in a controlled

temperature room at 25°C. Every 20 minutes, up to 1 h, the dripped volume was

recorded. The time (min) was plotted against the dripped volume (mL) and the slope was

taken as the melting rate.

4.2.3.3 Color
 The color analysis of the ice cream mix was performed by using Kinoca Minolta

CR-400 Chroma Meter. The colorimeter was calibrated using a white reference plate

prior to the ice cream measurement. The results were shown as the value of a*,b* and L*.

The final hue and chroma of the ice cream mix were calculated from the a* and b*.

4.2.3.4 pH measurement

The pH was measured from the melted ice cream sample using a pH meter model:

HANNA HI 98107.

4.2.3.5 Moisture content

Samples were weighed approximately 3-5 grams, and were analyzed by the rapid

moisture analyzer (Mettler Toledo HD43-S).

4.2.4 Protein analysis

Protein content in the ice cream was measured and defined by using the Kjeldahl method.

The protein factor used for calculation is 6.25.

4.2.4 Sensory Evaluation

The sensory evaluation was done by using the 9-point hedonic scale ranging from

extremely like to extremely dislike for several characteristics, such as texture, color, flavor,

sweetness, and overall acceptability. The sensory evaluation will be performed by 25-30

panelists who can consume whey proteins.

4.2.5 Statistic Analytical

SPSS was used to analyse the results performed during the sensory evaluation. Paired

t-test was as well used and the significant difference is defined at P<0.05.
 CHAPTER V

RESULTS AND DISCUSSION

Dairy ice cream is a complex food matrix composed of ice, air, fat, and the concentrated

liquid serum. Commonly, the ice cream characteristics are dependent on the formulation and its

processing conditions. The critical steps in manufacturing ice cream products are preparation of

the ice cream mix, homogenization, pasteurization, cooling, ageing, freezing, and churning. The

ice cream formulation and the stability of the ice cream emulsion can play a significant role in

determining the quality of the final product.

Table 5.1 : The paired t-test of two formulations of whey protein ice cream
Paired Differences

95% Confidence
Mean Std. Std. Interval of the t df Sig
Deviatio Error Difference (2-tailed)
n Mean
Lower Upper

Appearance -0.933 1.874 0.342 -1.633 -0.233 -2.728 29 0.011

Whey25%-Whey15%

Color -0.967 1.608 0.294 -1.567 -0.366 -3.293 29 0.003

Whey25%-Whey15%

Mouthfeel -1.000 2.017 0.368 -1.753 -0.246 -2.715 29 0.011

Whey25%-Whey15%

Aroma -0.467 2.029 0.371 -1.225 0.291 -1.259 29 0.218

Whey25%-Whey15%

Sweetness -0.933 2.180 0.398 -1.748 -.0119 -2.345 29 0.026

Whey25%-Whey15%

Flavour -0.833 2.397 0.437 -1.727 0.061 -1.907 29 0.66

Whey25%-Whey15%

Overall Liking -1.000 2.213 -0.404 -1.826 -0.173 -.2475 29 0.19

Whey25%-Whey15%

The objective of this study is to formulate an ice cream product with additional whey

protein for the athletes or physically-active individuals. Initially, two variations of whey protein

concentration were aimed at 15% and 25%. After the in-house sensory was performed, as shown

in Table 5.1, 15% whey protein substitution was selected as the best formulation of ice cream.

Moreover, 50% of low-fat milk was used to substituted half of the cream content in order to

reduce the fat in the product. The selected formulation was based on the results of the sensory

evaluation from 30 healthy panelists, age ranges from 18-22 years old. The result has shown that

the critical attributes which are the overall liking, aroma, and flavor are not statistically different.

15% Whey substitution was selected as a final formulation since high amount of whey protein

greatly impacts the color and mouthfeel of the ice cream. As illustrated in table 5.2 and table

5.3, it clearly signifies that 15% whey substitution receives the higher scores in both attributes,

color and mouthfeel. High amount of whey protein can possibly lead to overly-thick texture of

the ice cream due to the thickening effect of the protein (Young, 2007), thus reduces the sensory

scores. Moreover, the color of the ice cream is changed according to the amount of whey protein

added. 25% of whey substitution displayed yellow color in a darker shade than 15% whey

substitution. This as well might contribute to the lower scores given from the panelists.

Table 5.2: The paired samples statistics for color comparison between two formulations.
Mean N Std. Correlation Std. Error Mean

Color: Whey25% 6.700 30 1.534 0.280

Whey15% 7.667 30 0.959 0.175

Table 5.3: The paired samples statistics for mouthfeel comparison between two formulations.
 Mean N Std. Correlation Std. Error Mean

Mouthfeel: Whey25% 6.033 30 1.671 0.305

Whey15% 7.033 30 1.066 0.195

After the statistical analysis was done, the best ice cream formulation was finalized and

the overall composition of the ice cream was shown in Table 5.4 compared to the control sample

which did not contain any whey proteins and was partially substituted with low-fat milk.

Table 5.4 : The composition of each treatment of the ice cream.

Ingredients Control (g) Whey Ice cream

Milk 288 -

Half and Half Cream 128 64

Low-fat Milk - 64

Whey Protein Concentrate - 43.2

Water - 244.8

Stevia - 0.02

Sugar 80 50

Vanilla extract 2 (tbsp) 2 (tbsp)

 Figure 5.5 : Melting rate of the control sample and whey protein ice cream

Table 5.6 : The mean value of chemical and physical properties of whey protein ice cream
compared to the control sample
Ice cream mix Overrun (%) Moisture pH Melting Rate
Content (%)

Control 11.715±0. 06
𝑎
55.21±0. 15
𝑎
6.65±0. 07
𝑎
0.9069±0. 006
𝑎

Whey Protein 18.395±0. 91

𝑏
64.54±0. 22
𝑏
6.45±0. 07
𝑎
0.5252±0. 005
𝑏

Ice cream
𝑎,𝑏
Means in the same column without common superscripts differ (P<0.05).

The overall physical and chemical properties of the ice cream were illustrated in the

Table 5.6. The percent overrun of the ice cream is created by incorporating air into the ice cream

mix. This can influence the final quality of the ice cream because overrun can affect the hardness

of the ice cream(Muse & Hartel, 2004). As seen from the table, the sample with whey protein

powder exhibited an increase in % overrun. This raise could probably be a result of greater

foaming capacity of the whey protein powder which led to enhanced air bubbles. This resulted in
an excess volume of the ice cream (Akesowan, 2009). In addition, whey protein concentrate can

increase the overrun since the milk protein powder is subject to more significant total heat load

during manufacture because of the application of drying step. The whey protein powder was

more unfolded. This degree of denaturation can cause the proteins to absorb onto the air interface

faster than the protein content in regular liquid milk. It has been found that denatured proteins

have better foaming properties as a consequence of increased hydrophobicity and greater

interfacial contact (Alvarez et al., 2005). In addition, the formation of small ice crystals, can also

result from the increase in overrun. Patel et al (2006) have found that ice cream with additional

protein content tends to have smaller ice crystals because whey proteins and CN have good

water-binding capacity, therefore, free water is reduced and unable to form ice crystals.

Moreover, proteins can adsorb onto the surface of ice crystals and prevent the further growth of

larger ice crystals (Patel et al, 2006).

From the results, whey protein ice cream yields a higher amount of moisture content than

the control sample, 64.70 and 55.10 respectively. Giri et al. (2014) have explained that the

moisture content could possibly increase due to the replacement of sugar with stevia powder

which is a result of an increase in free moisture content.

For the pH measurement, the values of both samples were slightly different. However, it

did not differ statistically. This slight change in pH was in accordance with the study of Patel et

al (2006), it has stated that the titratable acidity in milk can be influenced by CN, albumin,

phosphates, citrates, and carbon dioxide. The decrease in pH could have resulted from the higher

amount of protein content in ice cream mix because the CN and albumin amount was raised

(Patel et al, 2006).

 According to the Table 5.6, Comparing to the control sample, the melting rate of whey

protein ice cream has decreased significantly. Several factors can cause the reduction in melting

rate of the ice cream. From the research, Mayangsari (2019) has observed that the substitution of

sugar with stevia leaf powder can highly affect the ice cream viscosity which leads to the melting

resistance. In stevia leaf powder, it is found to contain a fiber content. The fiber content can

increase the overall total solid content of the final ice cream product, as a consequence, the ice

cream becomes more viscous.The increased melting time of whey protein ice cream can also be

supported by the findings of Syed et al (2018) stating that protein content can improve the

melting property of the ice cream due to the fact that proteins adhere at the surface of water and

fat molecules. It causes the agglomeration of fat, resulting in more stabilized air cells. The

decrease in melting rate of the ice cream is also in accordance with the study of Rolon et al.

(2017) explaining that the less amount of fat can cause a reduction in melting rate due to the

replacement with low fat milk. The study has found that the samples with the highest amount of

fat exhibited an increase in particle size and a higher degree of fat destabilization resulting from

the higher probability of droplet collision at higher fat contents. Adapa et al. (2000) have also

observed a similar finding which the decrease in fat content could lead to the reduced rate in fat

destabilization.

Table 5.7 Values of the colorimetric parameters L*, a*, b*, hue and chroma of whey protein ice
cream compared to the control sample

Ice cream L* a* b* Hue Chroma

mix

Control 83.96±0. 15
𝑎
-2.66±0. 06
𝑎
12.67±0. 17
𝑎 78.14 12.94

Whey protein 75.8±0. 25

𝑏
0.61±0. 04
𝑏
20.45±0.04
𝑏 88.29 20.46
ice cream
 𝑎,𝑏
Means in the same column without common superscripts differ (P<0.05).

For the color determination of the ice cream mix, hue and chroma were calculated from

a* and b*. Hue angle was determined by using the formula: arctan(b*/a*). It refers to the degree

to which a stimulus can be described as similar to or different from stimuli that are described as

2 2
red, green, blue and yellow. While, the chroma was calculated from (𝑎 * ) + (𝑏 * ) .

Chroma is defined as the vividness of color. The color will become paler when the chroma is

reduced.The parameters of the color system as shown in Table 5.4 represent specific values. L*

represents the lightness which varies from black to white. a* specifies the variation of the color

from redness to greenness. Lastly, B* represents the variation of the color from yellowness to

blueness. The results shown in the table were in accordance with the study of Akesowan (2009),

which the sample with protein concentrate addition exhibited a lower L* values. The lower L*

values indicated that the color of the sample was darker than the control. The protein addition

could increase amine compounds, as a consequence, participated in Maillard reaction reacting

with the aldehydes and resulted in brown pigment formation. Values for a* and b* also increased

significantly compared to the control sample. The increase in these two values could signify that

the whey protein samples were more red and yellow color. The difference in color can be

explained by the color of the main ingredients. The control samples contain only regular milk

and half and half cream which appeared to be white color, on the other hand, the whey protein

ice cream contains whey protein powder which initially appears in light yellow.

Table 5.8: The nutritional composition of two ice cream samples with a serving size of 100 g

Ice cream mix Energy (Kcal) Fat (g) Protein (g) Carbohydrate (g)

Control 189.25 11.13 2.36 19.91

 Whey protein ice 133.8 5.98 8.22 12.81
cream

The overall nutritional composition of two ice cream samples were shown in Table 5.8.

There is an excellent reduction in total energy due to substitution in half and half cream and

sugar content. Fat and carbohydrates were decreased significantly while protein content was

increased by approximately 5.86%. The increase in protein can be due to the addition of whey

protein concentrate powder incorporated during the processing step. According to this table, the

ice cream could be claimed as reduced-fat ice cream. Prindiville et al. (1999) have specified ice

cream products into different categories depending on the amount of fat and any ice cream

products that contain 6%-9% fat content can be categorized into reduced-fat category. Moreover,

it can be claimed as a reduced-sugar ice cream. According to Thai RDI, any products contain

25% or higher reduction in overall sugar content can be labeled as reduced-sugar products. In

this study, the sugar was reduced approximately 37%, thus, it is able to be specified as the

reduced-sugar product. However, to be claimed as “high’’ protein, the product must contain

protein content more than 10 grams per 100 grams of food due to the regulations stated in the

Notification of the Ministry of Health. Therefore, this ice cream product shall be labeled as

“good source” of proteins which indicates that it contains more than 5 grams of protein per 100

grams of food. The future research should increase protein content if aims to be labeled as “high”

protein food. However, the changes in physical and chemical properties should be taken into

consideration as well.

CHAPTER VI
 CONCLUSION

The ice cream was successfully formulated using stevia powder as a partial substitution

for sugar. The low-fat milk and whey protein concentrate were fortified in order to cope with the

nutritional requirement for athletes or physically active individuals. The protein content was

increased by 5.86 g per 100 g. While the fat content and carbohydrates were reduced

significantly by 5.15% and 7.1%, respectively because of the replacement with low-fat milk and

stevia powder. The change in physical and chemical properties was observed. Overall, the

whey-enriched ice cream displays better characteristics in terms of overrun, melting rate, and

size of the ice crystals due to the increased protein content which contains higher amounts of

albumin and CN. It can only be claimed as reduced-fat and reduced-sugar ice cream due to the

fact that the protein content still does not meet the regulations required by the Ministry of Health

to be labeled as “high” protein food.

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 APPENDICES

APPENDIX A

Physical characteristics of vanilla ice cream with whey protein concentrate and vanilla ice

cream without whey protein concentrate (control)

Table A-A-1 : Percentage of overrun.

Sample %Overrun 1 %Overrun 2

With whey protein concentrate 18.46 18.33

Control 11.67 11.72

Table A-A-2 : Moisture Content.

Sample MC 1 MC 2

With whey protein concentrate 64.70 64.38

Control 55.10 55.32

Table A-A-3 : pH value

Sample pH 1 pH 2

With whey protein concentrate 6.4 6.5

Control 6.6 6.7

Table A-A-4 : Melting rate of ice cream.

Sample Melting Rate 1 Melting Rate 2

With whey protein concentrate 0.5195 0.5309

Control 0.9025 0.9113

Table A-A-5 : Colorimeter values of ice cream with whey protein concentrate.

Values Color 1 Color 2

L* 75.62 75.98

a* 0.58 0.64

b* 20.42 20.48

Table A-A-6 : Colorimeter values of ice cream without whey protein concentrate.

Values Color 1 Color 2

L* 83.86 84.07

a* -2.71 -2.62

b* 12.55 12.79
 APPENDIX B

Paired t-test from sensory evaluation using SPSS program.

Table A-B-1 : Paired samples statistics of sensory attributes between ice cream with 15%

whey protein concentrate and 25% whey protein concentrate.

Mean N Std. Deviation Std. Error Mean

Pair 1 APP15% 7.10 30 1.30 0.24

APP25% 6.17 30 1.72 0.31
Pair 2 Color15% 7.67 30 0.96 0.18
Color25% 6.70 30 1.53 0.28
Pair 3 MF15% 7.03 30 1.07 0.31
MF25% 6.03 30 1.67 0.19
Pair 4 Aroma15% 6.67 30 1.42 0.26
Aroma25% 6.20 30 1.70 0.31
Pair 5 Sweet15% 7.10 30 1.24 0.23
Sweet25% 6.17 30 1.53 0.28
Pair 6 Flavor15% 7.30 30 1.18 0.22
Flavor25% 6.47 30 1.72 0.31
Pair7 Overall15% 7.33 30 1.18 0.22
Overall25% 6.33 30 1.67 0.30

Table A-B-2 : Paired samples correlations of sensory attributes between ice cream with

15% whey protein concentrate and 25% whey protein concentrate.

N Correlation Sig

Pair 1 APP15%-APP25% 30 0.255 0.174

Pair 2 Color15%-Color25% 30 0.234 0.213
Pair 3 MF15%-MF25% 30 -0.039 0.836
Pair 4 Aroma15%-Aroma25% 30 0.158 0.405
Pair 5 Sweet15%-Sweet25% 30 -0.026 0.229
Pair 6 Flavor15%-Flavor25% 30 -0.344 0.063
Pair7 Overall15%- Overall25% 30 -0.180 0.340
 BIOGRAPHY

NAME Sarankorn Pornpetchlerk

DATE OF BIRTH 29/03/1997

PLACE OF BIRTH Bangkok, Thailand

INSTITUTIONS ATTENDED Thai-Chinese International School

Mahidol University International College

HOME ADDRESS 13/286 Rimkhlongbangkhor Road, Bangkhor,

Chomthong, Bangkok 10150
Tel. 086-515-5520
E-mail: sarankorn0329@gmail.com